Abstract: Synergy occurs when combined agents induce a response greater than the sum of their individual effects. The present invention provides new high throughput screening methods to detect agents acting synergistically in orthogonally pooled mixtures. Computational de-convolution of the pooled data with software reveals single-actives in the pools with twice the statistical power and with much greater efficiency than common high throughput screening approaches. Cross-correlating the orthogonal data reveals pools with activity that cannot be ascribed to any single compound. The components of such ‘Orphan’ activity pools are then tested individually and in all possible combination-pairs to identify and confirm synergy.